Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4371

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Roller Culture of Postimplantation Embryos

Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer

This protocol was adapted from "Isolation, Culture, and Manipulation of Postimplantation Embryos," Chapter 5 (Protocol 7, procedure provided by Patrick Tam), in Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

The first 100 words of the full text of this article appear below.


INTRODUCTION

This protocol describes roller culture of postimplantation embryos. By this method, embryos from 6.5 to 11.5 days post coitum (dpc) can be isolated, manipulated and cultured outside the uterus. Normal development in vitro will occur for 48 hours when embryos up to early somite stages are used, and for 24 to 36 hours when older embryos are used.


MATERIALS

Reagents

Dulbecco’s modified Eagle’s medium (DMEM)

Gas mixture (see table in Step 5)

Mouse embryos (6.5-11.5 dpc) (see Isolating Postimplantation Embryos: Early Primitive-Streak-Stage, Isolating Postimplantation Embryos: Late Primitive-Streak-Stage, Isolating Postimplantation Embryos: Early Neural-Fold-Stage, Isolating Postimplantation Embryos: Early Somite-Stage, depending . . . [Full Text of this Article]

Equipment


METHOD


Troubleshooting


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